Purification and characterization of histone H1 variants from human placenta
(2008) In Biotechnology and Bioprocess Engineering 13. p.772-777- Abstract
- Three histone H1 variants were extracted from human placental tissue in a single process using a high-salt buffer solution, and purified by ion exchange, hydroxyapatite, and reversed-phase chromatography. In the first chromatographic step, a cation exchanger resin, SP-Sepharose FF, was used to remove impurities having molecular weights higher than those of histones. In the second chromatographic step, hydroxyapatite resin was used to remove impurities with relatively low molecular weights. A second round of cation exchange chromatography using high-grade HS POROS resin resulted in two main fractions, each of which appeared as a single band following SDS-PAGE. The first fraction showed a single peak in RP-HPLC, while the second fraction... (More)
- Three histone H1 variants were extracted from human placental tissue in a single process using a high-salt buffer solution, and purified by ion exchange, hydroxyapatite, and reversed-phase chromatography. In the first chromatographic step, a cation exchanger resin, SP-Sepharose FF, was used to remove impurities having molecular weights higher than those of histones. In the second chromatographic step, hydroxyapatite resin was used to remove impurities with relatively low molecular weights. A second round of cation exchange chromatography using high-grade HS POROS resin resulted in two main fractions, each of which appeared as a single band following SDS-PAGE. The first fraction showed a single peak in RP-HPLC, while the second fraction showed two main peaks. These three peaks were further separated and polished by semi-preparative RP-HPLC, and their molecular masses and sequences were determined using MALDI-TOF-MS and N-terminal amino acid sequencing, respectively. The sequences and masses of these three variants corresponded with those of histones H1.2, H1.4, and H1.5. Moreover, all three purified histone subtypes demonstrated cytotoxicity in an MTT assay. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/83449036-979b-40a4-8bc6-a992ac1cc656
- author
- Pyo, Sang-Hyun
LU
; Lee, Jae-Heung
; Lee, Yeune Hee
; Yoon, Jong-Won
and Kim, Jin-Hyun
- publishing date
- 2008-12
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Biotechnology and Bioprocess Engineering
- volume
- 13
- pages
- 772 - 777
- publisher
- Korean Society for Biotechnology and Bioengineering
- external identifiers
-
- scopus:59849096865
- ISSN
- 1226-8372
- DOI
- 10.1007/s12257-008-0186-1
- language
- English
- LU publication?
- no
- id
- 83449036-979b-40a4-8bc6-a992ac1cc656
- date added to LUP
- 2025-09-19 11:26:10
- date last changed
- 2025-10-14 09:43:39
@article{83449036-979b-40a4-8bc6-a992ac1cc656,
abstract = {{Three histone H1 variants were extracted from human placental tissue in a single process using a high-salt buffer solution, and purified by ion exchange, hydroxyapatite, and reversed-phase chromatography. In the first chromatographic step, a cation exchanger resin, SP-Sepharose FF, was used to remove impurities having molecular weights higher than those of histones. In the second chromatographic step, hydroxyapatite resin was used to remove impurities with relatively low molecular weights. A second round of cation exchange chromatography using high-grade HS POROS resin resulted in two main fractions, each of which appeared as a single band following SDS-PAGE. The first fraction showed a single peak in RP-HPLC, while the second fraction showed two main peaks. These three peaks were further separated and polished by semi-preparative RP-HPLC, and their molecular masses and sequences were determined using MALDI-TOF-MS and N-terminal amino acid sequencing, respectively. The sequences and masses of these three variants corresponded with those of histones H1.2, H1.4, and H1.5. Moreover, all three purified histone subtypes demonstrated cytotoxicity in an MTT assay.}},
author = {{Pyo, Sang-Hyun and Lee, Jae-Heung and Lee, Yeune Hee and Yoon, Jong-Won and Kim, Jin-Hyun}},
issn = {{1226-8372}},
language = {{eng}},
pages = {{772--777}},
publisher = {{Korean Society for Biotechnology and Bioengineering}},
series = {{Biotechnology and Bioprocess Engineering}},
title = {{Purification and characterization of histone H1 variants from human placenta}},
url = {{http://dx.doi.org/10.1007/s12257-008-0186-1}},
doi = {{10.1007/s12257-008-0186-1}},
volume = {{13}},
year = {{2008}},
}